CF45-1, a secreted protein which participates in Dictyostelium group size regulation

Debra A. Brock, R. Diane Hatton, Dan Victor Giurgiutiu, Brenton Scott, Wonhee Jang, Robin Ammann, Richard H. Gomer

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Developing Dictyostelium cells aggregate to form fruiting bodies containing typically 2 × 104 cells. To prevent the formation of an excessively large fruiting body, streams of aggregating cells break up into groups if there are too many cells. The breakup is regulated by a secreted complex of polypeptides called counting factor (CF). Countin and CF50 are two of the components of CF. Disrupting the expression of either of these proteins results in cells secreting very little detectable CF activity, and as a result, aggregation streams remain intact and form large fruiting bodies, which invariably collapse. We find that disrupting the gene encoding a third protein present in crude CF, CF45-1, also results in the formation of large groups when cells are grown with bacteria on agar plates and then starve. However, unlike countin- and cf50- cells, cf45-1- cells sometimes form smaller groups than wild-type cells when the cells are starved on filter pads. The predicted amino acid sequence of CF45-1 has some similarity to that of lysozyme, but recombinant CF45-1 has no detectable lysozyme activity. In the exudates from starved cells, CF45-1 is present in a ∼450-kDa fraction that also contains countin and CF50, suggesting that it is part of a complex. Recombinant CF45-1 decreases group size in colonies of cf45-1 - cells with a 50% effective concentration (EC50) of ∼8 ng/ml and in colonies of wild-type and cf50- cells with an EC50 of ∼40 ng/ml. Like countin- and cf50- cells, cf45-1- cells have high levels of cytosolic glucose, high cell-cell adhesion, and low cell motility. Together, the data suggest that CF45-1 participates in group size regulation in Dictyostelium.

Original languageEnglish (US)
Pages (from-to)788-797
Number of pages10
JournalEukaryotic Cell
Volume2
Issue number4
DOIs
StatePublished - Aug 1 2003

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Dictyostelium
Proteins
Muramidase
Exudates and Transudates
Cell Adhesion

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

CF45-1, a secreted protein which participates in Dictyostelium group size regulation. / Brock, Debra A.; Hatton, R. Diane; Giurgiutiu, Dan Victor; Scott, Brenton; Jang, Wonhee; Ammann, Robin; Gomer, Richard H.

In: Eukaryotic Cell, Vol. 2, No. 4, 01.08.2003, p. 788-797.

Research output: Contribution to journalArticle

Brock, Debra A. ; Hatton, R. Diane ; Giurgiutiu, Dan Victor ; Scott, Brenton ; Jang, Wonhee ; Ammann, Robin ; Gomer, Richard H. / CF45-1, a secreted protein which participates in Dictyostelium group size regulation. In: Eukaryotic Cell. 2003 ; Vol. 2, No. 4. pp. 788-797.
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abstract = "Developing Dictyostelium cells aggregate to form fruiting bodies containing typically 2 × 104 cells. To prevent the formation of an excessively large fruiting body, streams of aggregating cells break up into groups if there are too many cells. The breakup is regulated by a secreted complex of polypeptides called counting factor (CF). Countin and CF50 are two of the components of CF. Disrupting the expression of either of these proteins results in cells secreting very little detectable CF activity, and as a result, aggregation streams remain intact and form large fruiting bodies, which invariably collapse. We find that disrupting the gene encoding a third protein present in crude CF, CF45-1, also results in the formation of large groups when cells are grown with bacteria on agar plates and then starve. However, unlike countin- and cf50- cells, cf45-1- cells sometimes form smaller groups than wild-type cells when the cells are starved on filter pads. The predicted amino acid sequence of CF45-1 has some similarity to that of lysozyme, but recombinant CF45-1 has no detectable lysozyme activity. In the exudates from starved cells, CF45-1 is present in a ∼450-kDa fraction that also contains countin and CF50, suggesting that it is part of a complex. Recombinant CF45-1 decreases group size in colonies of cf45-1 - cells with a 50{\%} effective concentration (EC50) of ∼8 ng/ml and in colonies of wild-type and cf50- cells with an EC50 of ∼40 ng/ml. Like countin- and cf50- cells, cf45-1- cells have high levels of cytosolic glucose, high cell-cell adhesion, and low cell motility. Together, the data suggest that CF45-1 participates in group size regulation in Dictyostelium.",
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